Investigation on different small RNA expression of TT19 suppressors under anthocyanin induction condition (AIC) using Small RNA-seq

To investigate the functions of TRANSPARENT TESTA 19 (TT19) on anthocyanin accumulation and stabilization using forward genetic screening approach. EMS mutagenized tt19 mutant to screen tt19 mutant suppressors that could restore the anthocyanin accumulation and the causative mutations were identified as point mutations in the RDR6/SGS3/DCL4 siRNA biogenesis pathway using whole genome re-sequencing. The idea for RNA-Seq and small RNA-Seq analysis is to illustrate the connection between siRNA biogenesis, TT19, and anthocyanin restoration. RNA's of 4 Arabidopsis lines (col-0, tt19 and two TT19 suppressors, all in triplicate) which were under AIC (3% sucrose shaking four days under 24h light) for four days was extracted. Small RNA libraries were prepared by BGI company and sequenced using BGISEQ-500 as 50SE. The RNA of 12 samples from 4 Arabidopsis lines (col-0, tt19 and two TT19 suppressors,all in triplicate) was extracted from seedlings which had been under AIC for four days. An analysis of the small RNAs differential expression among col-0, tt19, and TT19 suppressors was performed.